Method for reducing allergenicity in indoor spaces

ABSTRACT

According to this invention, allergenicity in an indoor space is reduced by denaturing allergens in allergen reservoirs that are capable of producing respiratory or skin reactions and physically removing the allergens from the allergen reservoirs in the indoor space. The allergen reservoirs may also be treated with pesticides and fungicides/fungistats to prevent reinfestation of house dust mites, molds and cockroaches.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of Ser. No. 11/529,586 filedSep. 29, 2006, which is a continuation-in-part of Ser. No. 11/239,473,filed Sep. 30, 2005, which is a continuation-in-part of U.S. applicationSer. No. 10/153,612, filed May 24, 2002, which claims the benefit ofU.S. Provisional Application No. 60/293,183, filed May 25, 2001. Theaforementioned applications are hereby incorporated by reference intheir entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to medically sound procedures for reduction inenvironmental allergenicity due to certain airborne allergens known toproduce asthma, allergic rhinitis and atopic eczema in both humans andanimals. Specifically, allergens derived from house dust mites, certainmolds, cockroaches, cats and dogs are removed from indoor living spacesto provide sanctuaries for asthma sufferers.

2. Review of Related Art

Asthmatic bronchitis (asthma), allergic rhinitis (hay fever), and atopiceczema are immunologic reactions which, in the majority of cases, aretriggered by one or more allergens from five primary sources: catdander, dog dander, mold spores, cockroach fecal pellets and house dustmite fecal pellets. All of the primary asthmatic allergens from thesefive sources are protein in nature, typically glycoprotein. Theseallergens accumulate on various indoor surfaces and in allergenreservoirs, such as carpets, upholstery, mattresses and pillows. Thepresence of these allergens in any interior space can make that spacehazardous for an asthmatic patient.

Of the primary allergens, house dust mite (HDM) accounts for 50-55% ofthe asthmatic's problems. HDMs proliferate indoors when temperature,humidity and the presence of sloughed epithelial cells (the mite'sprimary food source) are adequate for their survival and reproduction.Adult HDMs produce 20-30 fecal pellets per day per mite over a life spanof 2½ months. Each fecal pellet is heavily inoculated during its midgutformation with proteins (principally digestive enzymes) capable ofevoking an immune response.

HDM allergenic proteins have been identified for mites of the genusDermatophagoides, and the proteins fall mainly into two immunologicalgroups: Der I (Der pI and Der fI) and Der II (Der pII and Der fII). Theprimary sequence of HDM allergens has been disclosed (see a series ofU.S. patents and also reviews by Thomas, et al. (1998), “House-dust-miteallergens,” Allergy, 1998 September; 53(9):821-32; Platts-Mills T A, etal. (1997), “Indoor allergens and asthma: report of the ThirdInternational Workshop,” J Allergy Clin Immunol 1997 December; 100(6 Pt1):S2-24; and Chua K Y, et al. (1996), “Analysis of sequencepolymorphism of a major mite allergen, Der p 2,” Clin Exp Allergy. 1996July; 26(7):829-37). HDMs are primarily found in indoor textiles(carpet, upholstery, mattresses and pillows) where their population canbecome enormous—well into the millions. Mechanical disturbance canrelease the proteins from dried fecal pellets into the air, producing anallergenic cloud that will resettle onto textile surfaces over time.

Mold spores account for 10-15% of asthmatic reactions, and four moldgenera account for nearly all of the allergenic spores. These generaare: Alternaria, Cladiosporium, Aspergillus, and Penicillium, withAlternaria accounting for about 80% of the mold spore responses ofasthmatics. Cat dander accounts for 10-15%, cockroaches account for5-10%, and dog dander accounts for 5-10% of asthmatic reactions.

HDM, cockroach and mold allergens tend to be larger and heavier;therefore, they often settle on textile-covered surfaces, such ascarpets and upholstery. Even after being disrupted, such as when aperson walks on a carpet or sits on a sofa, the allergens will only riseup a few feet in the air (e.g., about three feet) and quickly settleagain.

Cat and dog allergens, on the other hand, tend to be smaller andlighter; thus, they often “float” in the air for up to six months.Eventually, those allergens will attach to a surface, such as atextile-covered surface, wall or ceiling. However, once they aredisrupted, the allergens will again “float” in the air for a lengthyperiod of time before settling.

Physicians currently address the disorders produced by allergens withsymptomatic medicinal approaches, desensitization methods and preventivemeasures such as textile removal from the home, air filters,dehumidifiers, mattress covers and the like, yet the problem stillexists. Ablative HDM chemicals currently in use include short terminsecticides or denaturants that reduce the allergenicity of proteinallergens, such as the proteins in HDM fecal pellets. Insecticides thathave been tested include benzyl benzoate (U.S. Pat. Nos. 5,916,917;6,107,341; and 6,117,440) phenyl salicylate, the organophosphate,pirimiphos methyl (Mitchell, et al., 1985, “Reduction of house dust miteallergen levels in the home: uses of the acaricide, pirimiphos methyl,”Clin. Allergy, 15:234), and synthetic pyrethroids such as permethrin(U.S. Pat. Nos. 5,843,981; 5,916,580; and 5,965,602; Glass, et al.,“Evaluation of the acaricide permethrin against all stages of theAmerican house dust mite Dermatophagoides farinae Hughes(Pyroglyphidae),” in Mitchell, et al., eds., Acarology IX Proceedings,Columbus, Ohio Biological Survey, 1997, pp. 693-695). InternationalPublication No. WO 98/30236 discloses a pesticide using protease enzymesto kill insects for the purpose of decreasing or eliminating theincidence of allergic reaction to dust, but this reference does notaddress allergic reactions to the enzyme. Tannic acid has been used todenature HDM fecal pellets short term (Green, “Abolition of allergens bytannic acid,” Lancet, 2:160 1984; U.S. Pat. No. 4,977,142), andpolyphenol denaturants have been used in combination with insecticides(Green, et al., 1989, “Reduction of house dust mites and mite allergens:Effects of spraying carpets and blankets with Allersearch DMS, anacaricide combined with an allergen reducing agent,” Clin. Exp. Allergy,19:203; U.S. Pat. No. 4,806,526).

Either the insecticide treatments or the chemical denaturants must bereapplied at 2-3 month intervals for any hope of efficacy. Theseproducts are available over-the-counter, and are subject to thevariability existing among do-it-yourselfers who apply such products.These products are also displaced from textiles during vacuuming andbecome air-borne, are inhaled, and can serve as a major irritant toasthmatics or hay fever sufferers. Therefore, there remains a need forimproved methods of reducing allergenicity in indoor spaces to alleviatethe suffering of asthmatics and hay fever sufferers.

SUMMARY OF THE INVENTION

It is an object of this invention to provide more effective methods formedically addressing asthma, allergic rhinitis and atopic eczema bycreating “sanctuary” spaces with low allergenicity.

This invention provides methods for reducing allergenicity in an indoorspace. In one embodiment, the method uses at least two of the followingsteps. One step provides for physically removing allergens capable ofproducing respiratory or skin reactions in asthmatics from allergenreservoirs in the indoor space. A second step provides for denaturingthe allergens in the allergen reservoirs to reduce allergic reaction tothe allergens. A third step provides for prophylactically preventingreinfestation of the indoor space by house dust mites and molds. In oneexemplary embodiment, the step of physically removing allergen fromallergen reservoirs is accomplished by vacuuming and hot water washing,the step of denaturing allergens is accomplished by enzymatic hydrolysisof the allergens and/or the step of preventing reinfestation isaccomplished by treating allergen reservoirs with a pesticide plus afungicide. In another exemplary embodiment, these steps are repeatedperiodically to maintain low levels of allergenicity.

In another embodiment, this invention provides a method for reducingallergenicity in an interior space comprising exposing textile-coveredsurfaces in an interior space to means for hydrolytically cleaving aplurality of peptide bonds in an allergenic polypeptide selected fromthe group consisting of house dust mite fecal antigens, cockroach fecalantigens, cat dander, dog dander and mold spores at ambient temperature,wherein the amount of active allergen in the interior space isconsequently reduced. In one exemplary embodiment, said means forhydrolytically cleaving peptide bonds is a subtilisin-type protease,more preferably, the subtilisin-type protease is subtilisin savinase.

In yet another embodiment, this invention provides a method for reducingallergenicity in an interior space comprising treating textile-coveredsurfaces in the interior space with one or more proteolytic enzymes,wherein said one or more proteolytic enzymes cleave peptide bonds inallergenic polypeptides selected from the group consisting of house dustmite fecal antigens, cockroach fecal antigens, cat dander, dog danderand mold spores, and wherein allergenicity in the interior space islowered. Typically, the textile-covered surface comprises a reservoirfor the allergenic polypeptides, and the reservoir may comprise one ormore of carpets, bedding, pillows or upholstery. Also, typically, thetextile-covered surface is not immersed in water in the method of thisinvention and/or the textile-covered surface is incubated with theproteolytic enzyme under ambient conditions. In an exemplary embodiment,the proteolytic enzyme is applied to said textile-covered surfaceperiodically, for example, semi-annually. In a another exemplaryembodiment, the method of this invention further comprises at least onestep selected from the group consisting of (a) vacuuming at least aportion of the allergenic polypeptide from the textile-covered surfaceand (b) washing the textile-covered surface with hot water to remove theallergenic polypeptide. In yet another exemplary embodiment, the one ormore proteolytic enzymes are applied to said textile-covered surface inconjunction with a pesticide that limits house dust mite infestation,such as permethrin. In still another exemplary embodiment, the one ormore proteolytic enzymes are subtilisin-type proteases. In even anotherexemplary embodiment, the enzyme is catalytically active at neutral pH.

In still another embodiment, this invention provides a method forreducing allergen load on an allergen reservoir comprising applying oneor more proteolytic enzymes to the reservoir, wherein the one or moreproteolytic enzymes cleave sufficient peptide bonds in allergenicpolypeptides selected from the group consisting of house dust mite fecalantigens, cockroach fecal antigens, cat dander, dog dander and moldspores to reduce active allergen load in the reservoir. In oneembodiment, the reservoir comprises animal fur and the one or moreproteolytic enzymes are dispensed from a porous glove. Typically, themethod of this invention is not carried out in a controlled reactionenvironment, but rather, the reaction conditions comprise ambienttemperature. In a preferred embodiment, the one or more proteolyticenzymes are applied in conjunction with a pesticide. In an exemplaryembodiment, the one or more proteolytic enzymes comprise asubtilisin-type protease, such as one that is catalytically active at aneutral pH. In another exemplary embodiment, the one or more proteolyticenzymes are applied contemporaneously with a pesticide and a fungicide.In a related embodiment, this invention provides anallergenicity-reducing composition comprising one or more proteolyticenzyme, a pesticide active against dust mites and a fungicide activeagainst Alternaria.

In an additional embodiment, the method comprises denaturing, inallergen reservoirs, allergens that are capable of producing respiratoryor skin reactions and physically removing the allergens from theallergen reservoirs in the indoor space. In one exemplary embodiment,the step of denaturing the allergens is accomplished by enzymatichydrolysis of the allergens, the step of physically removing theallergens from the allergen reservoirs is accomplished by hot waterwashing and, optionally, vacuuming. In another exemplary embodiment, themethod further comprises drying the allergen reservoir after the hotwater washing. In yet another exemplary embodiment, the enzymatichydrolysis is carried out by at least one proteolytic enzyme, optionallyby two or more proteolytic enzymes. In still another exemplaryembodiment, the at least one proteolytic enzyme is a subtilisin-typeprotease. The proteolytic enzyme may be incubated in ambient temperaturefor 1 to 30 minutes, such as 5 to 25 minutes or 10 to 20 minutes. Theproteolytic enzyme may be catalytically active at neutral pH.

In any of the aforementioned embodiments, the method may furthercomprise deactivating the proteolytic enzyme, such as with an enzymedenaturant. In addition, the method may further comprise limiting themobility, stabilizing, or physically restricting the movement of theenzyme in the environment through the addition of an adhesive. Enzymedenaturants that may be used include acetic acid, formic acid, sodiumhypochlorite, hydrogen peroxide and/or citric acid, particularly citricacid. The methods may further comprise prophylactically preventingreinfestation of the indoor space by house dust mites and molds, such asby treating the allergen reservoirs with a pesticide, for examplepermethrin, and a fungicide. Moreover, in any of the aforementionedembodiments, the at least one proteolytic enzyme is delivered with anagglomerating agent, such as polyester, acrylic latex, polyvinylacetate, solubilized starch, and polyvinyl alcohol (particularlypolyvinyl alcohol). The allergens to be denatured in any of theembodiments are selected from the group consisting of house dust mitefecal antigens, cockroach fecal antigens, cat dander, dog dander andmold spores. The methods of the invention may be repeated periodically,such as annually. The allergen reservoirs of the aforementionedembodiments comprise textile-covered surfaces, such as carpets, bedding,pillows and upholstery, which may or may not be immersed in water.

In another embodiment of the invention, a method for reducingallergenicity in an indoor space comprises aerosolizing at least oneproteolytic enzyme, optionally two or more proteolytic enzymes, in theindoor space, wherein said at least one proteolytic enzyme cleavessufficient peptide bonds in allergenic polypeptides selected from thegroup consisting of house dust mite fecal antigens, cockroach fecalantigens, cat dander, dog dander and mold spores to reduce activeallergen load in the indoor space. In an exemplary embodiment, the atlease one proteolytic enzyme is delivered with an agglomerating agent,for example, film-forming agents or glues, such as polyesterdispersions, acrylic latex, polyvinyl acetate, solubilized starch, andpolyvinyl alcohol, particularly polyvinyl alcohol. In another exemplaryembodiment, the method may further comprise deactivating the proteolyticenzyme, such as with an enzyme denaturant, or limiting the mobility ofthe enzyme through the addition of an adhesive. Enzyme denaturants thatmay be used included acetic acid, formic acid, sodium hypochlorite,hydrogen peroxide and citric acid, particularly citric acid. In yetanother exemplary embodiment, the method further comprises physicallyremoving the allergenic polypeptides from allergen reservoirs in theindoor space by hot water washing, and optionally, vacuuming. The methodmay further comprise drying the allergen reservoirs after the hot waterwashing. In still another exemplary embodiment, the method furthercomprises prophylactically preventing reinfestation of the indoor spaceby house dust mites and molds, such as by treating the allergenreservoirs with a pesticide, for example permethrin, and a fungicide. Inany of the aforementioned methods, the allergen reservoirs are incubatedwith the at least one proteolytic enzyme for 1 to 30 minutes, such as 5to 25 minutes and 10 to 20 minutes. The at least one proteolytic enzymemay comprise a subtilisin-type protease and be catalytically active atneutral pH.

In one additional embodiment, a method for A method for reducingallergenicity in an indoor space comprises: applying at least oneproteolytic enzyme to allergen reservoirs; incubating the at least oneproteolytic enzyme at ambient temperature for 1 to 30 minutes;deactivating the proteolytic enzyme; physically removing the allergensfrom the allergen reservoirs; and treating the allergen reservoirs witha pesticide and a fungicide.

In another embodiment, this invention provides a method for reducingallergenicity in an interior space comprising exposing textile-coveredsurfaces in an interior space to negatively charged polymers such as thefilm-forming polymers or glues described above. Such polymers includepolyester dispersions, acrylic latex, polyvinyl acetate, solubilizedstarch, and polyvinyl alcohol, particularly polyvinyl alcohol.Additionally, the negatively charged polymers include natural andsynthetic absorbent polymers, e.g., natural cellulosic polymers such ascarboxymethylcellulose nitrate and synthetic polymer such as acrylicacid polymers. Preferably, the polymers are applied in conjunction withan antimicrobial or antifungal agent.

In another embodiment, this invention provides a method for reducingallergenicity in an interior space comprising coating or impregnating anair filter matrix with negatively charged polymers and allowing the airto pass through the coated or impregnated air filter matrix, thusremoving the allergens from the air. The polymers include thefilm-forming polymers or glues described above and include polyesterdispersions, acrylic latex, polyvinyl acetate, solubilized starch, andpolyvinyl alcohol, particularly polyvinyl alcohol. Additionally, thenegatively charged polymers include natural and synthetic absorbentpolymers, e.g., natural cellulosic polymers such ascarboxymethylcellulose nitrate and synthetic polymer such as acrylicacid polymers. In this embodiment, the coated or impregnated matrix maybe periodically moistened so as to maintain the anionic nature of thepolymers. Preferably, the polymers are applied in conjunction with anantimicrobial or antifungal agent.

In another embodiment of the invention, a method for reducingallergenicity in an indoor space comprises aerosolizing negativelycharged polymers, such as the film-forming polymers or glues describedabove. Such polymers include polyester dispersions, acrylic latex,polyvinyl acetate, solubilized starch, and polyvinyl alcohol,particularly polyvinyl alcohol. Additionally, the negatively chargedpolymers include natural and synthetic absorbent polymers, e.g., naturalcellulosic polymers such as carboxymethylcellulose nitrate and syntheticpolymer such as acrylic acid polymers. Preferably, the polymers areapplied in conjunction with an antimicrobial or antifungal agent.

In yet another exemplary embodiment, the method further comprisesphysically removing the allergen from allergen reservoirs in the indoorspace by hot water washing, and optionally, vacuuming. The method mayfurther comprise drying the allergen reservoirs after the hot waterwashing.

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

This invention relates to medical and veterinary medicine proceduresaddressing certain allergens known to produce asthma, allergic rhinitisand atopic eczema in both humans and animals. The specific allergens arederived from house dust mites, certain molds, cockroaches, cats anddogs. These allergens are proteins, typically complex glycoproteins,having polypeptide structures which render the molecule allergenic.

The goal of this invention is to offer the immunologically compromisedpatient an organized, professional and holistic approach to control themajor indoor antigens (derived from HDM, molds, cockroach, cat and dog)that are capable of producing the medical conditions asthmaticbronchitis, allergic rhinitis and atopic eczema. The major habitat forthese antigens is textiles which provide adequate humidity, temperatureand food for proliferation of the sources of these allergenic antigens.Antigen prevention should address temperature, humidity and food, buttaken individually or collectively, simply addressing these factors inany combination is grossly inadequate to decrease antigen levels belowestablished medical statistical antigen levels capable of incitingallergic response in a sensitized individual.

Within the indoor environment, the primary asthmatic allergens arecommonly found in allergen reservoirs, such as textiles (carpet,upholstery, mattresses, pillows, etc.). According to this invention,allergenicity in an indoor space is reduced by denaturing allergens inallergen reservoirs that are capable of producing respiratory or skinreactions and physically removing the allergens from the allergenreservoirs in the indoor space. The allergen reservoirs may also betreated with pesticides and fungicides/fungistats to preventreinfestation of house dust mites, molds and cockroaches.

DEFINITIONS

For the purpose of describing the present invention, the following termsare defined.

“Indoor space” means an enclosed space containing a body of air forwhich passage of air into and out of the space is hindered (e.g., bysolid walls, semi-permeable screens, doors, windows, etc.), so thatparticulates or aerosols suspended in the atmosphere of the indoor spacewill dissipate only slowly by settling or exchange with the atmospherein adjacent spaces through limited connecting passages (e.g., throughdoors and windows or by diffusion through screens). Examples of indoorspaces include the interior of a residential dwelling, a room or suiteof rooms that is not subject to free exchange of air with the outdoors,the inside of airplanes, buses, automobiles, trains, hotel rooms,offices, assembly or meeting halls, etc.

“Primary allergens” as discussed herein are allergens found in catdander, dog dander, mold spores, cockroach fecal pellets and house dustmite fecal pellets that are associated with asthma, allergic rhinitisand atopic eczema in both humans and animals.

“Allergen reservoir” means any region or volume in an indoor space thatcan dispense allergens when mechanically disturbed. Typical reservoirsare woven materials including textiles such as carpets, upholstery,pillows and bedding or baskets and mats. Allergens may accumulate in thereservoirs by settling from an allergen aerosol or by deposition from aninfestation of insects of other sources of allergens.

“Active allergens” are chemical entities, typically macromolecules, thatcause an allergic reaction in a sensitized individual. If the chemicalentities are chemically modified so that they no longer cause theallergic reaction, the entities have become inactive allergens. Theallergenic effect is typically caused by binding of regions on themacromolecular allergen to receptors in the sensitized individual. Theseregions are called epitopes, and modification of the epitopes can resultin inactive allergens.

“Allergen load” means the level of active allergen in some volume orarea, which may be an indoor space or an allergen reservoir. Typically,allergen load is measured by collecting the allergen in some volume orarea (e.g., by filtering allergen from an aerosol or by mechanicallydisturbing an allergen reservoir and collecting the allergen released)and measuring the amount of allergen, for example by ELISA.

To “lower allergenicity” means reducing the allergen load in thedesignated space or reservoir. Target allergen level for house dust miteallergen is less than 2 micrograms/gram dust; levels above 10micrograms/gram dust are likely to produce some symptoms inmite-sensitive asthmatics, and 100 micrograms/gram dust is likely toprovoke an asthma attack. While it is preferred to reduce HDM allergenlevels to below 100 micrograms/gram dust or lower, any reduction of theallergen level is beneficial to allergy sufferers. Indeed, the amount ofrelief an allergy sufferer experiences is directly proportional to thereduction in allergen level. Thus, reducing the allergen level by 10%may provide approximately 10% of relief. Although a 100% reduction inallergenicity is optimum, a reduction of allergenicity in a designatedspace or reservoir of at least 30% is a significant benefit to allergysufferers.

“Ambient conditions” refers to conditions that will affect the rate ofchemical reactions, such as temperature, pressure, etc., as thoseconditions exist normally in contrast to controlled conditions whichoccur inside closed vessels and consciously deviate from ambient tofacilitate some action. Ambient conditions exist, for example, in abedroom or a garden, while controlled conditions exist inside arefrigerator or a washing machine or a chemical reactor.

Proteases are enzymes which catalyze the hydrolysis of polypeptides toproduce peptide fragments. The polypeptides consist of various differentamino acid residues connected by peptide bonds, and proteases maycatalyze hydrolysis of bonds between certain amino acid residues inpreference to other peptide bonds. “Subtilisin-type proteases” arepeptide-hydrolyzing enzymes equivalent to the proteases secreted byvarious species of Bacillus. The proteases from Bacillus species arerelatively non-selective among the various peptide bonds. In particular,subtilisin savinase, derived from Bacillus lentus, and subtilisin BPN,derived from B. amyloliquefaciens have been shown to be effective.

Physical Removal of Allergens

Studies have shown that dry vacuum can remove up to 50-55% of the housedust mite material from allergen reservoirs, such as carpets.Effectiveness of the vacuum protocol may be confirmed by ELISA (see,e.g., Example 2, below). As an example, a portion of carpet might bydivided into two test plots, and each extracted with hot water, afterone of the plots had been vacuumed. The effectiveness of the vacuum maybe determined by comparing the amount of HDM substance in the twoextracts by ELISA. Procedures for measuring the allergen level in anenvironmental area are also disclosed in U.S. Pat. No. 5,679,535, whichis incorporated herein by reference, and the skilled artisan will beable to develop variant methods from the description therein.

Even more effective allergen removal from allergen reservoirs can beachieved by hot water washing, such as extraction. The hot watertypically used on an allergen reservoir is from 120° F. to 160° F.,depending on the equipment used. In one embodiment, the hot waterincludes surfactants to facilitate mobilizing the allergenic materialfrom the textile fibers. Such a procedure has been shown to removesubstantially all of the allergens (<2 microgram of the house dust mitesubstance/gram dust). Effectiveness of the hot water wash and/orextraction may also be confirmed by ELISA as described above.

In one exemplary embodiment of the invention, water at a temperature of120-140° F. is applied to a textile-covered surface, such as a carpet,followed promptly by vacuum removal of as much of the water as possible(typically 95%). In another exemplary embodiment, the textile-coveredsurface is washed and then, after removal of most of the water, thesurface is dried with a dryer, such as a high-power industrial fan.Removal of water and/or drying is important for the prevention of moldovergrowth.

The hot water wash may occur before, during or after the degradation ofallergens, or any combination thereof. For example, the allergenreservoir may be washed, and then a solution comprising the allergendenaturant, such as a hydrolyzing enzyme, may be applied. In anotherexample, the allergen denaturant may be in the wash solution. As anotherexample, the allergen reservoir may be washed after degradation of theallergens, and the wash solution may comprise an enzyme deactivator.

Enzymatic Degradation of Allergens

As discussed above, the primary asthmatic allergens are glycoproteins.Proteases (which may be genetically engineered) are capable ofcatalyzing the hydrolytic cleavage of these listed allergenicglycoproteins into peptide fragments, thereby denaturing theirallergenicity. The proteolytic fragments are both less allergenic andmore easily mobilized from the textile during the physical removalprocess. Therefore treatment with enzymes further reduces theallergenicity associated with treated textiles. Surprisingly, thefragments do not form highly allergenic aerosols.

Any enzyme that will cleave the allergenic polypeptides to destroy theepitopes recognized by the asthmatic's sensitized immune system issuitable, but proteases that are less specific (i.e., hydrolyze peptidebonds between a variety of different amino acid residues, rather thanonly between specific residues) are preferred. In particular,subtilisin-type proteases from various Bacillus sp. have been shown tobe effective, including subtilisin savinase, derived from Bacilluslentus, or subtilisin BPN, derived from B. amyloliquefaciens. Similarnon-specific proteases are suitable for use in this invention, as wellas Bacillus neutral proteases and the like. Genetically engineeredenzymes are within the contemplation of this invention, so long as theresulting enzyme can be shown to destroy allergenic epitopes of theprimary allergens, as shown by ELISA tests (see, e.g., U.S. Pat. No.5,314,991). Suitable assays may be analogous to the tests for active Tcell epitopes of Der pI, Der pII, Der fI, and Der fII disclosed in U.S.Pat. No. 5,968,526 (incorporated herein by reference).

Application of the enzymes to the textile may be by any suitableprocedure. The enzymes may be applied in a separate step, with the hotwater used for physical removal, or with the prophylacticpesticide/fungicide discussed below. Typical formulations will bebuffered to avoid extremes of pH that are detrimental to enzymestability, and may include other well known enzyme stabilizers, such aspolyols, calcium salts or other ionic stabilizers.

Since allergens may become airborne when the allergen reservoir isdisturbed and some may even “float” for a lengthy period of time, it isadvantageous, in some cases, to deliver the enzyme with an agglomeratingagent. When the enzyme is aerosolized, agglomeration occurs due to theconcentration of solutes as the water evaporates. The agglomeratingagent will not only help to attract an allergen to the enzyme, but itwill also help to weigh down the allergen so that it will settle out ofthe air more quickly. Moreover, the large size of the enzyme/agglomerantmolecule will help inhibit inhalation into the lungs. Agglomeratingagents that may be used include polyester, acrylic latex, polyvinylacetate, solubilized starch or polyvinyl alcohol (PVA).

In a particular mode, an aqueous solution of the enzyme is absorbed in aporous matrix that is then run over the allergen reservoir to betreated, and the enzyme solution is dispensed into the reservoir bycapillary action. One example of such a porous matrix is a sheet ofsponge-like material incorporated into the palm of a glove that is wornby a person when petting a cat or dog. The enzyme solution is applied tothe animal's coat from the sponge, and the solution will penetrate thecoat to the animal's skin (adjacent to sebaceous glands) to degrade allor a portion of the allergenic antigen in the fur reservoir (especiallycat dander antigen). This method is particularly useful as part of theveterinary treatment of asthmatic pets discussed in more detail below.

In another embodiment of the invention, the enzyme solution is deliveredvia aerosol or fogger (collectively referred to as “aerosolizedenzyme”). The enzyme solution may be aerosolized through use of, e.g., atotal release canister or a spray gun. The aerosolized enzyme wouldallow the enzyme to come in to contact with the smaller and lighterallergens, such as those from cat and dog dander, that are “floating” inthe air. In one exemplary embodiment, the enzyme is delivered with anagglomerating agent as described above so that the allergen will “stick”to the enzyme/agglomerant molecule. This will not only provide for theattraction of the allergen to the enzyme, but it will also provide theneeded weight for the allergen to settle out of the air.

In another exemplary embodiment, the aerosolized enzyme is negativelycharged, such as through an electrostatic filter or other methods knownin the art, so that the enzyme molecule will attract allergens, whichare positively charged. The aerosolized enzyme may optionally containstabilizing agents to enhance durability (e.g. of the enzyme or theaerosol). In one embodiment, the negatively charged aerosolized enzymeis delivered with an agglomerating agent as previously described.

Controlling the Allergenicity of the Degrading Enzyme

It is well known that high concentrations of enzymes may cause allergicreactions. Therefore, it is recommended that the enzyme concentration onan indoor surface be less than 65 μg/m².

To address this issue, one embodiment of the invention provides a methodthat further comprises deactivating the enzyme after hydrolysis hasoccurred. In one exemplary embodiment, the enzyme is deactivated byusing an enzyme denaturing agent, such as acetic acid, formic acid,sodium hypochlorite, hydrogen peroxide or citric acid. In an exemplaryembodiment, the enzyme denaturant is an acidic solution, such as0.5-5.0% citric acid solution.

In one embodiment of the invention, the enzyme denaturant is appliedbefore the physical removal of the allergen. In another embodiment, theenzyme denaturant is applied with the hot water washing solution used tophysically remove the allergen. In yet another embodiment, the enzymedenaturant is applied after the physical removal of substantial portionsof the allergen.

In an exemplary embodiment of the invention, at least one proteolyticenzyme, such as a subtilisin-type protease, is applied to the allergenreservoir; incubated at ambient temperature for 1 to 30 minutes, forinstance 10-20 minutes, and deactivated with an enzyme denaturant, suchas an acidic solution. Then, the allergens are physically removed thefrom the allergen reservoir, for example, by hot water washing, which isfollowed by drying. Finally, the allergen reservoir is treatedprophylactically, such as with a pesticide and a fungicide as describedbelow.

Another embodiment that may be used to prevent allergic reactions to theproteolytic enzyme comprises the addition of an adhesive to theformulation containing the enzyme, so that when the composition isapplied and dries, the adhesive adheres the enzyme to the appliedsurface. When the enzyme is adhered to the surface, allergic reactionsto the enzyme may be prevented as the enzyme is no longer able to becomeairborne and is in effect sequestered on the surface. The adhesives area subset of the aforementioned agglomerating agents that are capable ofimparting adhesion and include polyester such as polyester dispersions,acrylic latex, polyvinyl alcohol, polyvinyl acetate, starch orcombinations thereof. The adhesives may also be selected to impart adesired degree of flexibility when dried. The particular adhesivepolymers useful in the present embodiment are known to one of ordinaryskill in the art and may be selected to impart the desired degree ofadhesiveness and flexibility described above.

According to a preferred embodiment, the adhesive acts as a binder andin the liquid state allows the enzyme to agglomerate, thus weighing downthe enzyme. As the treatment dries, the adhesive anchors the enzyme tothe surface.

When the enzyme is applied in combination with an adhesive, the adhesiveimparts a degree of safety to the composition in that the enzyme is nolonger free to move about within the environment. Because of the reducedmobility of the enzyme, allergic reactions that may result due to thepresence of the enzyme are reduced or eliminated.

The enzyme and adhesive may be applied to any surface including, but notlimited to: textiles, such as carpet; filters, such as air filters; orany other surface that requires treatment against allergens.

In another embodiment, the enzyme may be used in a waste treatmentcomposition, for example to treat odors in clay (e.g., kitty litter). Inthis embodiment, the adhesive adheres the enzyme to clay in the drystate. When the clay is wetted, the enzyme is released, exhibiting thedesired waste treatment activity. Upon drying, the enzyme is re-anchoredto the clay surface. The enzyme and adhesive may be added to claycompositions in a manner known to one of ordinary skill in the art, suchas the methods set forth in U.S. Pat. Nos. 5,634,431 and 6,207,143,which are herein incorporated by reference in their entirety.

Agglomeration of Allergen Through Treatment with Anionic Polymers

The negatively charged polymers will attract allergens that arepositively charged. The polymers therefore act to remove such allergensfrom the environment through agglomeration of the allergen and polymeras a result of this ionic interaction. The allergen/polymer agglomeratedcomplexes that are formed can then be removed from the treated area asdescribed below or allowed to remain in the treated area, with theallergen sequestered in the complex. Suitable negatively chargedpolymers include polyester, acrylic latex, polyvinyl acetate,solubilized starch or polyvinyl alcohol (PVA). As described above, suchnegatively charged polymers are preferably adhesive in nature, and thuswill bond to surfaces to which they are applied, e.g., textiles and airfilter matrices. Additionally, the negatively charged polymers includenatural and synthetic absorbent polymers, e.g., natural cellulosicpolymers such as carboxymethylcellulose nitrate and synthetic polymersuch as acrylic acid polymers. Preferably, the polymers are applied inconjunction with an antimicrobial or antifungal agent. Applications ofthe negatively charged polymers to suitable surfaces are describedbelow.

Application of the negatively charged polymers to a textile may be byany suitable procedure. The negatively charged polymers may be appliedin a separate step, with the hot water used for physical removal, orwith the prophylactic pesticide/fungicide discussed below.

Coating or impregnating of the negatively charged polymer to an airfilter matrix may be made by any known procedure, including, e.g., spraycoating. In one such procedure, a polymer latex may be spray coated ontoand allowed to penetrate into a filter matrix. In order to maintain theionic nature of the polymers, the filter matrix typically isperiodically rewetted, e.g. through spraying the coated or impregnatedfilter matrix with water. Various air filtration media are well known,for example those in HVAC systems having matrices which include, but arenot limited to fiberglass and fibrous webs of addition polymers, e.g.,polypropylene fibers.

In a particular mode, an aqueous solution of the polymer is absorbed ina porous matrix that is then run over the allergen reservoir to betreated, and the polymer solution is dispensed into the reservoir bycapillary action. One example of such a porous matrix is a sheet ofsponge-like material incorporated into the palm of a glove that is wornby a person when petting a cat or dog. The polymer solution is appliedto the animal's coat from the sponge, and the solution will penetratethe coat to the animal's skin (adjacent to sebaceous glands). Thismethod is particularly useful as part of the veterinary treatment ofasthmatic pets discussed in more detail below.

In another embodiment of the invention, the polymer solution isdelivered via aerosol or fogger (collectively referred to as“aerosolized polymer”). The enzyme may be aerosolized through use of,e.g., a total release canister or a spray gun. The aerosolized polymerwould allow the polymer to come into contact with the smaller andlighter allergens, such as those from cat and dog dander, that are“floating” in the air. In one exemplary embodiment, the allergen will“stick” to the polymer molecule. This will provide the needed weight forthe allergen to settle out of the air, as well as allowing the allergento be sequestered in the complex as described above.

Prophylactic Treatment

House dust mites and mold spores are ecologically interlocked. Moldspores are the principal diet of immature dust mites. Furthermore, moldspores settle on sloughed epithelial cells which form the principal foodsource for adult dust mites, and mold secretes enzymes that predigestthe epithelial cell proteins to make them more easily assimilated by themites. Thus, reducing mold spore levels will have a limiting effect onmite infestation. Together, HDM and mold spores account for abouttwo-thirds of the asthmatic's problems, so restraining thereaccumulation of these two allergens will provide significant relieffor the asthma sufferer.

Application of a pesticide targeted at mites and a microbicide/stattargeted at the allergenic molds can maintain the low level ofallergenicity achieved by physical cleaning and/or enzymatic degradationof the allergens in a target area. Pesticides that have some effectagainst dust mites are known, and use of such pesticides is within thecontemplation of this invention. Pesticides that are effective againstboth house dust mites and cockroaches may be used, but even morepreferred are pesticides of extremely low human toxicity that areeffective against the major allergen-producer—house dust mites.

Permethrin is a synthetic pyrethroid that is less toxic to humans thanmost other pyrethroid derivatives. Permethrin is currently used fortreating head lice and scabies; an ointment containing 5% permethrin isapplied for 12 hours, then washed off. Less than 2% of the permethrin isabsorbed through the skin. Permethrin is a particularly preferredpesticide for use in the methods of this invention since it can achieve100% kill of house dust mites in carpet at levels of only 0.1%permethrin. Another class of suitable pesticides are polyheterocycliccompounds, such as ivermectin, avermectin or abamectin (collectivelyreferred to as “Avermectins”). Avermectins are typically applied tocontrol mites on foliage or fruit at about 0.014%. Avermectins may beused at concentrations of less than 0.014% for carpets.

As discussed above, application of pesticide is likely to be moreeffective if it is accompanied by an antimicrobial compound effective asa fungicide or fungistat. (The term “fungicide” will be used hereinafterto represent either fungicides or fungistats, or combinations thereof.)In particular, fungicides should be chosen for their effectivenessagainst the four mold genera responsible for asthma-inducing allergens:Alternaria, Cladiosporium, Aspergillus, and Penicillium, mostparticularly Alternaria. The skilled artisan will be aware of suitabletests for fungicidal efficacy, including inhibition of fungal growth inpetri dishes or shake flask cultures. Preferably, the antimicrobialcompound selected as fungicide according to this invention will bestable when formulated under conditions suitable for permethrin.Organophosphate 2-ethylhexyl esters (antimicrobial chemicals sold underthe trademark INTERSEPT) are suitable, as is asoxystrobin. Otherantimicrobials meeting the criteria provided herein may be selected bythe skilled worker.

Typically, the pesticide and fungicide are formulated in an aqueousmixture or in other volatile solvent(s) for application to the textilesthat might serve as allergen reservoirs. The mixture may be sprayed ontothe textiles or painted on or applied by any other method that providesa relatively even coating throughout the body of the textile.Application methods also include padding, sponging, foaming, e.g.,aerosol and spraying (e.g., power and pump). The solvent willvolatilize, leaving a residue of pesticide and fungicide to serve toinhibit development of new mite infestations and/or mold inoculation ofthe textile material. The presence of the pesticide/fungicide willpostpone the need for repeat cleaning to maintain low allergen load inthe indoor space occupied by the textiles. Preferably, the pesticide andthe fungicide are chosen to retain efficacy as applied for substantiallysimilar periods. Most preferably, both the pesticide and the fungicideas applied will retain efficacy for at least six months or even for overa year.

Prophylactic compositions containing permethrin and asoxystrobin are ofparticular interest because both of these agents migrate into textilefibers where they are ionically bound, resulting in extended periods ofefficacy and reduced volatility for lower toxicity and allergenicity ofthe prophylactic agents. Another desirable formulation is an aerosolthat can deposit active components in the same reservoir areas reachedby cat dander allergen, including light fixtures, ceiling fans, cabinettops, etc. Application of the prophylactic compositions in heating orair conditioning ducts, either as aerosols or by impregnating air filtermatrices in the ducts, is also within the contemplation of thisinvention.

Veterinary Treatment

Animals are also susceptible to allergic asthmatic reaction against thesame allergens that affect humans, particularly HDM. In particular,asthma has been diagnosed in household pets, such as cats and dogs.Allergen reservoirs associated with animal asthma include the animal'scoat, pet beds or sleeping mats, and litter boxes. In addition totreatment to apply enzymes to an animal's coat with a porous glove asdescribed above, animal asthma may be treated by applying enzymes to thetextiles in pet beds or a pesticide/fungicide combination to either petbed textiles or to the animal's coat. Enzymes and/or pesticide/fungicidecombinations may also be added to the pet's bath water or dispersed inan indoor space by aerosol bomb. To prevent allergic reactions (eitherin the pet or in the owner) from the enzyme, the enzyme may bedeactivated, for example with an enzyme denaturant. Alternatively, or inaddition to the enzyme deactivation, the allergen reservoir may bewashed with hot water.

EXAMPLES

In order to facilitate a more complete understanding of the invention,Examples are provided below. However, the scope of the invention is notlimited to specific embodiments disclosed in these Examples, which arefor purposes of illustration only.

Example 1 Allergen Control System

Major university laboratory and double-blinded clinical studies haveproven the following indoor textile “cradle to grave” holistic approachto be capable of maintaining HDM and HDM fecal pellet antigens belowlevels capable of producing allergenic response (2 microgram HDM antigenper gram of dust):

Phase One

Ideally carpet, upholstery and bedding should be anti-antigen treatedduring manufacturing bilaterally with SMITE™, i.e., syntheticpermethrin, and UNISEPT™ i.e., fungistat/fungicide¹. HDM antigen is themajor indoor antigen (55%) which coexists with mold antigen (10-15% ofindoor antigen load). There is synergism between HDMs and mold spores inthat:

-   -   a) Immature HDMs food source consists of mold spores    -   b) Mold spores predigest epithelial cells which is the major        food source of adult HDMs. ¹Phosphoric acid, bis(2-ethylhexyl)        ester, compd.

Accordingly new textile products for indoors that are involved in thisissue should be prophylactically controlled against both HDMs and mold.Fungistats/fungicides must be specific in addressing the four molds thatare predominate in creating immunological response (Alternaria 80%),Cladosporium, Penicillium, Aspergillus)

Phase Two

A. New products containing SMITE™ and UNISEPT™ deteriorate over timenecessitating re-application of the acaricide and fungicide at intervalsof one year. The textile product must be dry vacuumed, then subjected tohot water extraction for dirt, dust, HDMs, mold spores and otherantigens prior to applying the pesticides. Hot water extractionobviously leaves moisture residual which translates into mold overgrowthand proliferation, thereby necessitating application of UNISEPT™ alongwith the acaricide SMITE™.

B. In-Use Products (textiles)—mite/mold infested: Priority-wise, thehighest concentration of HDMs exists in mattresses and pillows followedby upholstery. The greatest numbers of HDMs exists in carpet. Thusly,in-use textiles that are a habitat for mites and mold should likewisebe:

-   -   a) Professionally vacuumed (dry) which will remove 50% of dust,        dirt, HDMs, mold spores and other antigens in carpet as proven        in our laboratories.    -   b) Followed by professionally hot water extraction which will        remove 100% of the HDM antigen, as laboratory tests have proven.    -   c) Followed by professional application of SMITE™ and UNISEPT™.        Professional applicators will apply these pesticides with a        special nozzle to insure:        -   Even distribution of the chemicals to textiles to control            mite and mold regrowth for one year after antigen removal        -   This specialized nozzle will also assure even in-depth            penetration of chemicals for HDMs, which, although blind,            abhor light and accordingly are embedded in textiles well            below the surface.    -   d) Professional drying procedures are practiced to inhibit mold        overgrowth.

These in-use textile procedures should be duplicated at approximatelyone year intervals for maximum antigen protection even though:

-   -   a) SMITE™ will control 85% of the HDM population for one year        before declining slowly to 52% HDM control and 48 months.    -   b) UNISEPT™ is near 100% effective against all immunologically        significant molds for one year.

Both SMITE™ and UNISEPT™ are EPA registered for these specific goals,are water miscible, user friendly toxicity-wise, basicallynondisplacable, low volatility, odorless, dye and fabric friendly.

Phase Three

The major antigens this invention addresses emanate from HDMs, moldspores, cockroaches, cat and dog. All of these antigens are allergenscomposed of glycoproteins at the molecular level. The antigenicity ofthe allergen is manifested via the amino acid radicals comprising thecomplex glycoprotein. The inventor has proven that the HDM antigen canbe completely denatured of all activity immunologically with the use ofgenetically engineered enzymes called proteases. Protease can bespecific for proteins, carbohydrates or fats in disassembling theirspecific molecules, or a combination of proteases can be used todenature and fragmentize complex glycoproteins. The resultant fragmentsare non-allergenic.

Application methodology of proteases to denature the antigens of HDMs,mold, cockroach, cat and dog herein discussed would include:

-   -   Protease use with detergents to clean textiles infested with        antigens.    -   Impregnate heating/AC filters with proteases.    -   Aerosol bombing of areas infested with antigens    -   Pet usage: Proteases on grooming tools, in bath water, in litter        boxes, in sleeping mats, etc.

Example 2 Measurement of Allergen Load

Allergen load in carpet is measured by vacuuming a measured square yardof carpet with a hand-held vacuum, having a dust collecting bag, bystroking for 30 seconds in each of two perpendicular directions. Theallergen content is determined by ELISA measurement normalized per gramof collected dust.

IMMULON™ microtiter plates are bound with monoclonal antibodies (mAb)specific for individual antigens. After washing plates withphosphate-buffered-saline (PBS) 2×, the plates are saturated with 100microliters of 1% bovine serum albumin in PBS. Next 100 microliter ofdoubling serial dilutions from 250 ng/mL to 0.5 ng/mL of referenceallergen standard of pre-set dilutions of samples are added and allowedto incubate for one hour. Following incubation, plates are washed 5×with PBS and wells are coated with biotinylated specific mAb and allowedto incubate, then washed and coated with streptavidin-peroxidasesolution. Bound biotinylated-labeled antibody is detected by using2′,2′-azino-bis(3-ethyl-benzthiazoline)sulfonic acid as a chromogen andH₂O₂ as a substrate. Each plate will be read when the optical density ofthe highest standard reaches 2.2 using a DYNATECH™ microplatespectrophotomer at 405 nm. Level of active allergen in each sample willbe determined by interpolation from the reference curves and reported inmicrogram antigen/gram dust.

Example 3 Efficacy of Allergen Denaturant on HDM, Cockroach, and DogAllergens

Eight different Allergen Denaturant solutions were prepared using fourdifferent subtilisin-type proteases and two different binding agents[AD1-AD4 had UNIBOND 1808, which contains water-dispersible polyesterresin, and AD5-AD8 had UNIBOND 1809, which contains solubilizedpolyvinyl alcohol (both binding agents are from Unichem, Inc., HawRiver, N.C.)]. Each solution was tested against a standard to ensurethat the Allergen Denaturant solutions did not interfere withcommercially available mAb ELISA tests (INDOOR Biotechnologies, Inc.,Charlottesville, Va.) used to measure exposure to allergens. Detectionwas performed by using a POWERWAVE 200 (Biotek Instruments, Inc.,Winooski, Vt.). The test showed that the Allergen Denaturant solutionsdid not interfere with the ELISA system.

The Allergen Denaturant solutions were then tested with Der p 2 miteallergen, Can f 1 dog allergen and Bla g 2 cockroach allergen. First,the ELISA plates were coated with anti-Der p 2 mAb 1D8, anti-Can f 1 mAb6E9 and anti Bla g 2 mAb 7C11 overnight at 4° C. The three allergenswere incubated with the eight Allergen Denaturant solutions and a PBScontrol, for a total of 27 samples, and incubated for two hours at roomtemperature.

After incubation, the samples were diluted 1/100, 1/200, 1/400 and 1/800and tested with the commercial ELISA kits and detected as describedabove. Curves of the four dilutions for each sample were generated andcompared to the standard curve. An average of the 4 dilutions, asdetermined by the standard curve, was calculated to determine theconcentration of allergen remaining after exposure to the AllergenDenaturant solutions. The effect of the Allergen Denaturant solutionswere expressed as a percent reduction of the allergen concentrationafter exposure to the solutions compared to the concentration of theallergen without an Allergen Denaturant solution (PBS Control). Resultsof the tests are shown below in Tables 1 to 3 below.

TABLE 1 Efficacy of Allergen Denaturant Solutions on Mite AllergenAllergen Denaturant Solution Der p 2 (μg/ml) % Reduction Der p 2 PBSControl 322.5 AD1 312.0 3 AD2 349.6 −8 AD3 347.0 −8 AD4 128.6 60 AD5300.0 7 AD6 382.0 −18 AD7 196.0 39 AD8 94.5 71

TABLE 2 Efficacy of Allergen Denaturant Solutions on Dog AllergenAllergen Denaturant Solution Can f 1 (μg/ml) % Reduction Can f 1 PBSControl 149.5 AD1 128.0 14 AD2 138.0 8 AD3 72.0 52 AD4 45.0 70 AD5 71.252 AD6 145.6 3 AD7 49.3 67 AD8 2.2 99

TABLE 3 Efficacy of Allergen Denaturant Solutions on Cockroach AllergenAllergen Denaturant Solution Bla g 2 (μg/ml) % Reduction Bla g 2 PBSControl 555.9 AD1 47.8 91 AD2 59.7 89 AD3 125.3 77 AD4 101.0 82 AD5272.3 51 AD6 433.0 22 AD7 252.3 55 AD8 144.5 74

The Allergen Denaturant solutions had significant effect on the Der p 2mite allergen, Can f 1 dog allergen and Bla g 2 cockroach allergencompared to the PBS Control. Moreover, AD4 and AD8 had a greater effecton the various antigens than the other enzyme solutions, except for AD1& AD2 with respect to the cockroach antigen.

Example 4 Efficacy of Allergen Denaturant on Mold Allergens

Three different Allergen Denaturant solutions were prepared using threedifferent subtilisin-type proteases and UNIBOND 1808 (AD9-AD11). Eachsolution was tested against a standard to ensure that the AllergenDenaturant solutions did not interfere with commercially available mAbELISA tests (INDOOR Biotechnologies, Inc., Charlottesville, Va.) used tomeasure exposure to allergens. Detection was performed by using aPOWERWAVE 200 (Biotek Instruments, Inc., Winooski, Vt.). The test showedthat the Allergen Denaturant solutions did not interfere with the ELISAsystem.

The Allergen Denaturant solutions were then tested with Asp f1 and Alt a1 mold allergens. First, the ELISA plates were coated with anti-Asp f 1mAb 4A6 and anti-Alt a 1 mAb 121 overnight at 4° C. The two allergenswere incubated with the six Allergen Denaturant solutions and a PBScontrol, for a total of 8 samples, and incubated for two hours at roomtemperature.

After incubation, the samples were diluted 1/100, 1/200, 1/400 and 1/800and tested with the commercial ELISA kits and detected as describedabove. Curves of the four dilutions for each sample were generated andcompared to the standard curve. An average of the 4 dilutions, asdetermined by the standard curve, was calculated to determine theconcentration of allergen remaining after exposure to the AllergenDenaturant solutions. The effect of the Allergen Denaturant solutionswere expressed as a percent reduction of the allergen concentrationafter exposure to the solutions compared to the concentration of theallergen without an Allergen Denaturant solution (PBS Control). Resultsof the tests are shown below in Tables 4 an 5 below.

TABLE 4 Efficacy of Allergen Denaturant Solutions on Asp f 1 MoldAllergen Allergen Denaturant Solution Asp f 1 (μg/ml) % Reduction Asp f1 PBS Control 149.9 AD9 100.3 33 AD10 91.4 39 AD11 135.6 10

TABLE 5 Efficacy of Allergen Denaturant Solutions on Alt a 1 MoldAllergen Allergen Denaturant Solution Alt a 1 (μg/ml) % Reduction Alt a1 PBS Control 231.8 AD9 143.3 38 AD10 151.2 35 AD11 77.3 67

The Allergen Denaturant solutions had moderate effects on the Asp f 1and Alt a 1 mold allergens, with AD11 having the greatest effect on Alta 1.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the appended claims. Modificationsof the above-described modes for carrying out the invention that areobvious to persons of skill in medicine, veterinary medicine, allergy &immunology, entomology, enzymology, pharmacology, and/or related fieldsare intended to be within the scope of the following claims.

All publications and patent applications mentioned in this specificationare indicative of the level of skill of those skilled in the art towhich this invention pertains. All such publications and patentapplications are herein incorporated by reference to the same extent asif each individual publication or patent application was specificallyand individually indicated to be incorporated by reference.

1.-56. (canceled)
 57. A method for reducing allergenicity in ambient airof an indoor space in need thereof, said ambient air having at least onepositively charged airborne allergen and said indoor space havingallergen reservoirs, said at least one positively charged airborneallergen having polypeptide structures that render the at least onepositively charged airborne allergen allergenic, said at least onepositively charged airborne allergen selected from the group consistingof house dust mite fecal antigens, cockroach fecal antigens, cat dander,dog dander, mold spores, and combinations thereof, the methodcomprising: a. aerosolizing a negatively charged polyester into ambientair of an indoor space in a manner that completely fills the indoorspace with the aerosolized polyester; b. the aerosolized polyesterstaying airborne for a period of time that will allow the aerosolizedpolyester to attract at least one positively charged airborne allergenand weigh down the at least one positively charged airborne allergen sothat the at least one positively charged airborne allergen will settleinto an allergen reservoir; and c. removing the at least one positivelycharged airborne allergen from the allergen reservoir.
 58. The method ofclaim 57 further comprising a step after step b. of: physically removingthe at least one positively charged airborne allergen from the allergenreservoirs in the indoor space by hot water washing.
 59. The method ofclaim 58 further comprising a step of: drying the allergen reservoirsafter the hot water washing.
 60. The method of claim 57 wherein: saidaerosolizing is accomplished via a fogger.
 61. A method for reducingallergenicity in ambient air of an indoor space in need thereof, saidambient air having at least one positively charged airborne allergen andsaid indoor space having allergen reservoirs, said at least onepositively charged airborne allergen having polypeptide structures thatrender the at least one positively charged airborne allergen allergenic,said at least one positively charged airborne allergen selected from thegroup consisting of house dust mite fecal antigens, cockroach fecalantigens, cat dander, dog dander, mold spores, and combinations thereof,the method comprising: a. aerosolizing a negatively chargedagglomerating agent into ambient air of an indoor space in a manner thatcompletely fills the indoor space with the aerosolized polyester; b. theaerosolized agglomerating agent staying airborne for a period of timethat will allow the aerosolized agglomerating agent to attract at leastone positively charged airborne allergen and weigh down the at least onepositively charged airborne allergen so that the at least one positivelycharged airborne allergen will settle into an allergen reservoir; and c.removing the at least one positively charged airborne allergen from theallergen reservoir.
 62. The method of claim 61 wherein: said negativelycharged agglomerating agent is an acrylic latex.
 63. The method of claim61 wherein: said negatively charged agglomerating agent is a polyvinylacetate.
 64. The method of claim 61 wherein: said negatively chargedagglomerating agent is a solubilized starch.
 65. The method of claim 61wherein: said negatively charged agglomerating agent is a polyvinylalcohol.
 66. The method of claim 61 wherein: said negatively chargedagglomerating agent is an acrylic latex.
 67. The method of claim 61further comprising a step after step b. of: physically removing the atleast one positively charged airborne allergen from the allergenreservoirs in the indoor space by hot water washing.
 68. The method ofclaim 67 further comprising a step of: drying the allergen reservoirsafter the hot water washing.
 69. The method of claim 61 wherein: saidaerosolizing is accomplished via a fogger.